Skin plays a crucial role in many important physiologic functions including fluid homeostasis, sensory detection, and protection against extemal insults. Damage to skin represents a major clinical problem, and can result from multiple events such as mechanical trauma, bums, and pathologic conditions (e.g. cancer and diabetes). Autografting of skin is the gold standard of treatment for skin repair, however the amount of donor skin is in limited supply, and the quality of donor skin can be compromised in certain patients. Allografts and xenografted tissues are often used in place of autografts, however these substrates also present disadvantages including the possibility of immune reactivity and transfer of pathogens. For all of these reasons, there is currently intensive interest in generating alternative materials that can serve as skin substitutes, or as temporary supports that induce skin regeneration. The principal goal of the proposed research is to develop a biodegradable fibroblast-embedded matrix that mimics the architecture, mechanical properties, biochemistry and cellularity of skin and that con-espondingly stimulates skin tissue regeneration. To achieve this objective, our laboratory has developed a novel layer-by-layer electrospinning method to create a nanofibrous matrix with dermal fibroblasts distributed throughout the scaffold thickness. The nanofibers are composed of a blend of collagen 1, a principal constituent of the dermis, with polycaprolactone (PCL), a synthetic polymer that imparts mechanical strength to the matrix, and allows tuning of scaffold degradation rate. Preliminary studies suggest that scaffolds generated by this approach support the survival and proliferation of embedded fibroblasts, and also stimulate keratinocytes grown on the top of the scaffolds to undergo differentiation and formation of a stratefied epidermal layer.